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Human TNF-α(Tumor Necrosis Factor Alpha) ELISA Kit
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Product No

FY-EH1405

Sample Type

Serum, Plasma, Tissue homogenate and Other biological samples；Sample Volume=100μL

Assay Time

3.5H

Price($)

280 400

Size

48T 96T

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Product Details
Product Name in English:
Human TNF-α(Tumor Necrosis Factor Alpha) ELISA Kit
Species:
Human
Alternative Name:
APC1 protein; Cachectin; Cachetin; DIF; TNF; TNF, monocyte-derived; TNFA; TNF-A; TNFalpha; TNF-alpha; TNF-alphacachectin; TNFATNF, macrophage-derived; TNFG1F; TNFSF1A; TNFSF2; TNFSF2TNF superfamily, member 2; tumor necrosis factor (TNF superfamily, member 2); tumor necrosis factor alpha; Tumor necrosis factor ligand superfamily member 2; tumor necrosis factor; tumor necrosis factor-alpha
Uniport ID:
P01375
Assay Type:
Sandwich
Standard:
200pg/mL
Sensitivity:
0.18pg/mL
Detection Range:
3.13-200pg/mL
Assay Time:
3.5H
Sample Type:
Serum, Plasma, Tissue homogenate and Other biological samples；Sample Volume=100μL
Test Principle:
The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to TNF-α. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to TNF-α. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain TNF-α, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of TNF-α in the samples is then determined by comparing the OD of the samples to the standard curve.
Technical Data
As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.
Concentration OD1 OD2 Average OD Corrected
200 2.347 2.364 2.356 2.284
100 1.553 1.56 1.557 1.49
50 0.922 0.93 0.926 0.859
25 0.439 0.442 0.441 0.376
12.5 0.26 0.262 0.261 0.197
6.25 0.16 0.161 0.161 0.097
3.13 0.113 0.114 0.114 0.05
0 0.063 0.063 0.063 0
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level were tested on 3 different plates, 20 replicates in each plate, respectively.
Item Intra-assay Precision Intra-assay Precision
Sample 3 3
Replicate 9 18
CV(%) 5 8
Rate of recovery
The recovery of spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
Sample Type Range(%) Average Recovery(%)
Serum (n=8) 90-105 97
EDTA plasma (n=8) 85-97 91
Cell culture media (n=8) 96-107 101
Linear
Samples were spiked with high concentrations of target proteins and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.
Serum (n=5) EDTA plasma (n=5) Cell culture media (n=5)
1:2 Range (%) 80-101 95-104 86-93
Average (%) 87 103 91
1:4 Range (%) 87-96 82-96 91-105
Average (%) 89 85 98
1:8 Range (%) 96-105 92-105 87-99
Average (%) 98 100 92
1:16 Range (%) 95-103 83-92 99-105
Average (%) 103 89 100
Assay Procedures
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